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目的 研究人外周血树突状细胞 (DendriticCell,DC)联合LPAK细胞体外诱导人肝癌细胞株BEL 74 0 2凋亡的作用 ,并进行形态学分析。方法 实验分LD、L、D和B四组 ,采用中性红摄入比色法检测细胞毒活性 ,原位末端标记 (TUNEL)技术、电镜技术观察细胞的光镜形态和超微结构。结果 D组与B组比较吸光度值无显著性差异 ,即无杀伤活性 (P >0 .0 5) ;LD组与L组比较 ,细胞毒活性为LD组 >L组 (P <0 .0 1)。光镜下凋亡的肿瘤细胞呈TUNEL阳性 ,细胞核染成棕黄色或深棕色 ,大小不等 ,形态也不一 ;电镜下肿瘤细胞核染色质凝聚、固缩和胞质浓缩 ,凋亡小体多见 ;少部分LPAK细胞内形成自噬体 ,发生自噬性凋亡。结论 人血DC联合LPAK细胞能有效地诱导肝癌细胞凋亡 ,在此过程中LPAK细胞同时发生自噬性凋亡。
Objective To study the effect of human peripheral blood dendritic cells (DCs) combined with LPAK cells on inducing apoptosis of human hepatocellular carcinoma cell line BEL 74 0 2 in vitro and perform morphological analysis. Methods The experiment was divided into four groups: LD, L, D and B. Neutral red colorimetric method was used to detect cytotoxicity. TUNEL technique and electron microscope were used to observe the morphology and ultrastructure of the cells. Results There was no significant difference in absorbance value between group D and group B, ie no killing activity (P > 0.05). Compared with group L, the cytotoxic activity of group LD was greater than that of group L > L (P < 0.01). ). Under light microscopy, the apoptotic tumor cells were TUNEL positive, and the nuclei were brownish or dark brown with different sizes and shapes. Under electron microscopy, the nuclear chromatin condensation, condensation and cytoplasm condensation, apoptotic bodies See; a small number of LPAK cells form autophagosomes and undergo autophagic apoptosis. Conclusion Human hematopoietic DC combined with LPAK cells can effectively induce apoptosis of hepatocellular carcinoma cells. During this process, autophagic apoptosis occurs simultaneously in LPAK cells.